IP Library › Granted Patent US 12,234,295
Granted Patent B2
US 12,234,295 · App. 17/815,110 · Granted Feb 25, 2025

Chimeric transmembrane protein comprising antibody dimerization domains and a type I cytokine receptor endodomain, encoding nucleic acids thereof and methods of use thereof

Inventors: Martin Pulé (London, GB); Shaun Cordoba (London, GB); Matteo Righi (London, GB); James Sillibourne (London, GB); Shimobi Onuoha (London, GB); Simon Thomas (London, GB)
Assignee: AUTOLUS LIMITED
C07K16/3069A61K39/001116A61K39/001135A61K39/001182A61K39/001194A61K39/4611A61K39/4631A61K39/464494A61K39/464495C07K14/46C07K14/7155C12N5/0636A61K2039/55522C07K2317/56C07K2317/622C07K2319/70
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,234,295
App. No.
17/815,110
Granted
Feb 25, 2025
Kind
B2
Abstract

The present invention provides a chimeric cytokine receptor (CCR) comprising: (i) an exodomain which binds to a ligand selected from a tumour secreted factor, a chemokine and a cell-surface antigen; and (ii) a cytokine receptor endodomain.

Claims (52)

1. A chimeric transmembrane protein comprising:

(i) a first polypeptide which comprises

(a) a first dimerization domain comprising the dimerization portion of an antibody heavy chain constant domain (C H ), and

(b) a first chain of a Type I cytokine receptor endodomain; and

(ii) a second polypeptide which comprises

(a) a second dimerization domain comprising the dimerization portion of an antibody light chain constant domain (C L ), which dimerizes with the first dimerization domain, and

(b) a second chain of a Type I cytokine receptor endodomain;

wherein the first and second polypeptides spontaneously dimerize.

2. The chimeric transmembrane protein according to claim 1 , wherein:

(i) the first polypeptide comprises

(a) a heavy chain constant domain (C H ), and

(b) a first chain of the Type I cytokine receptor endodomain; and

(ii) the second polypeptide comprises

(a) a light chain constant domain (C L ) and

(b) a second chain of the Type I cytokine receptor endodomain.

3. The chimeric transmembrane protein according to claim 2 , wherein the first polypeptide comprises an antibody heavy chain variable domain (V H ) and a heavy chain constant domain (C H ); and the second polypeptide comprises an antibody light chain variable domain (V L ) and a light chain constant domain (C L ).

4. A nucleic acid construct which comprises

(i) a first nucleic acid comprising a nucleic acid sequence encoding a first polypeptide which comprises:

(a) an antibody heavy chain constant domain (C H ) and

(b) a first chain of a Type I cytokine receptor endodomain; and

(ii) a second nucleic acid comprising a nucleic acid sequence encoding a second polypeptide which comprises:

(a) an antibody light chain constant domain (C L ) and

(b) a second chain of the Type I cytokine-receptor endodomain.

5. The nucleic acid construct according to claim 4 which also comprises a nucleic acid sequence encoding a chimeric antigen receptor (CAR), where the CAR comprises an antigen-binding domain, a spacer domain, a transmembrane domain, and an endodomain, wherein the antigen-binding domain specifically binds a target antigen.

6. A vector comprising a nucleic acid construct according to claim 4 .

7. A method for making a cell, which comprises a step of introducing: a nucleic acid that comprises a nucleic acid sequence that encodes the chimeric transmembrane protein of claim 1 into a cell.

8. A cell which comprises the chimeric transmembrane protein according to claim 1 .

9. The cell according to claim 8 , which also comprises a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen-binding domain, a spacer domain, a transmembrane domain, and an endodomain, wherein the antigen-binding domain specifically binds a target antigen.

10. A cell which comprises a chimeric transmembrane protein according to claim 2 .

11. The cell according to claim 10 , which also comprises a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen-binding domain, a spacer domain, a transmembrane domain, and an endodomain, wherein the antigen-binding domain specifically binds a tumour-associated antigen (TAA).

12. A pharmaceutical composition comprising a plurality of cells according to claim 8 .

13. A pharmaceutical composition comprising a plurality of cells according to claim 9 .

14. A pharmaceutical composition comprising a plurality of cells according to claim 10 .

15. A pharmaceutical composition comprising a plurality of cells according to claim 11 .

16. A method of killing a target cell in a subject, which comprises a step of administering a pharmaceutical composition according to claim 13 to the subject, wherein the target cell expresses the target antigen.

17. A method for treating a subject for cancer, which comprises the step of administering a pharmaceutical composition according to claim 13 to the subject, wherein the target antigen comprises a tumor-associated antigen (TAA) expressed at the surface of cells of the cancer.

18. The method according to claim 17 , which comprises:

(i) isolation of a cell-containing sample from a subject;

(ii) transduction or transfection of T cells or NK cells from the sample with (a) a nucleic acid construct or (b) a vector comprising the nucleic acid construct; said nucleic acid construct encoding the chimeric transmembrane protein and the CAR; and

(iii) formulating the cells from (ii) to make the pharmaceutical composition.

19. A method for treating a subject for cancer, which comprises the step of administering a pharmaceutical composition according to claim 15 to a subject.

20. A kit which comprises:

(i) a first vector comprising a nucleic acid sequence encoding a first polypeptide which comprises

(a) an antibody heavy chain constant domain (C H ), and

(b) a first chain of a Type I cytokine receptor endodomain; and

(ii) a second vector comprising a nucleic acid sequence encoding a second polypeptide which comprises

(a) an antibody light chain constant domain (C L ), and

(b) a second chain of a Type I cytokine receptor endodomain.

21. The kit according to claim 20 which also comprises a vector comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen-binding domain, a spacer domain, a transmembrane domain, and an endodomain, wherein the antigen-binding domain specifically binds a target antigen.

22. A kit which comprises:

i) a vector comprising the nucleic acid according to claim 4 , and

ii) a vector comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen-binding domain, a spacer domain, a transmembrane domain, and an endodomain, wherein the antigen-binding domain specifically binds a target antigen.

Assignments (2)
PATENT SECURITY AGREEMENT Recorded Jul 30, 2026
From: AUTOLUS LIMITED
To: PERCEPTIVE CREDIT HOLDINGS V, LP, AS ADMINISTRATIVE AGENT
Reel/Frame 076084/0283 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 9, 2022
From: PULE, MARTIN; CORDOBA, SHAUN; RIGHI, MATTEO; SILLIBOURNE, JAMES; ONUOHA, SHIMOBI; THOMAS, SIMON
To: AUTOLUS LIMITED
Reel/Frame 061704/0037 →
Priority Claims (1)
GB 1514875 · Aug 20, 2015 · national
Continuity (4)
Continuation 16998756 · Aug 20, 2020
Division 16113224 · Aug 27, 2018
Continuation 15753486
Related Publication 20230133682A1 · May 4, 2023
References Cited (59)
US 10800854B2 · Pule et al. · 2020 [cited by applicant]
US 10800855B2 · Pule et al. · 2020 [cited by applicant]
US 20130280220A1 · Ahmed et al. · 2013 [cited by applicant]
US 20180244797A1 · Pule et al. · 2018 [cited by applicant]
US 20180305433A1 · Pule et al. · 2018 [cited by applicant]
US 20180312570A1 · Pule et al. · 2018 [cited by applicant]
US 20200360432A1 · Pule et al. · 2020 [cited by applicant]
US 20210040227A1 · Pule et al. · 2021 [cited by applicant]
US 20210040228A1 · Pule et al. · 2021 [cited by applicant]
RU 2522004C2 · 2014 [cited by applicant]
WO WO9429458A1 · 1994 [cited by applicant]
WO WO2004029244A1 · 2004 [cited by applicant]
WO WO2007115230A2 · 2007 [cited by applicant]
WO WO2008045437A2 · 2008 [cited by applicant]
WO WO2009003145A1 · 2008 [cited by applicant]
WO WO2010085660A2 · 2010 [cited by applicant]
WO WO2012138858A1 · 2012 [cited by applicant]
WO WO2013123061A1 · 2013 [cited by applicant]
WO WO2015150771A1 · 2015 [cited by applicant]
WO WO2016061574A1 · 2016 [cited by applicant]
Kloss C.C., et al., “Combinatorial Antigen Recognition with Balanced Signaling Promotes Selective Tumor Eradication by Engineered T Cells,” Nature Biotechnology, New York, doi:10.1038/nbt.2459, ISSN 1087-0156, XP0551306… [cited by applicant]
Stuhlmann-Laeisz C., et al., “Forced Dimerization of gp130 Leads to Constitutive STAT3 Activation, Cytokine-Independent Growth, and Blockade of Differentiation of Embryonic Stem Cells,” Molecular Biology of the Cell, Ju… [cited by applicant]
Kloss et al., Combinatorial Antigen Recognition with Balanced Signaling Promotes Selective Tumor Eradication by Engineered T Cells, Nature Biotechnology, 31(1): 71-75, (Jan. 2013). [cited by applicant]
Stuhlmann-Laeisz et al., Forced Dimerization of gp130 Leads to Constitutive STAT3 Activation, Cytokine-independent Growth, and Blockade of Differentiation of Embryonic Stem Cells, Molecular Biology of the Cell, 17: 2986… [cited by applicant]
Abate-Daga et al., “A Novel Chimeric Antigen Receptor Against Prostate Stem Cell Antigen Mediates Tumor Destruction in a Humanized Mouse Model of Pancreatic Cancer,” Human Gene Therapy, 25(12):1003-1012 (2014). [cited by applicant]
Bayat et al., “Production and Characterization of Monoclonal Antibodies against Human Prostate Specific Antigen,” Avicenna J Med Biotechnol, 7(1):2-7 (2015). [cited by applicant]
Chang et al., “Five Different Anti-Prostate-specific Membrane Antigen (PSMA) Antibodies Confirm PSMA Expression in Tumor-associated Neovasculature,” Cancer Research 59:3192-3198 (1999). [cited by applicant]
Chinnasamy et al., “Local Delivery of Interleukin-12 Using T Cells Targeting VEGF Receptor-2 Eradicates Multiple Vascularized Tumors in Mice,” Clinical Cancer Research 18(6):1672-1683 (2012). [cited by applicant]
Communication pursuant to Article 94(3) in European Application No. 16 756 758.5 dated Apr. 29, 2020 (12 pages). [cited by applicant]
Donnelly et al., “The ‘cleavage’ activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring ‘2A-like’ sequences,” J Gen Virol, 82(5):1027-1041 (2001). [cited by applicant]
Guedan et al., “ICOS-based chimeric antigen receptors program bipolar TH17/TH1 cells,” Blood, 124(7):1070-1080 (2014). [cited by applicant]
Hassuneh et al., “Evidence for the Participation of Interleukin-2 (IL-2) and IL-4 in the Regulation of Autonomous Growth and Tumorigenesis of Transformed Cells of Lymphoid Origin,” Blood 89:610-620 (1997). [cited by applicant]
Hillerdal et al., “Systemic treatment with CAR-engineered T cells against PSCA delays subcutaneous tumor growth and prolongs survival of mice,” BMC Cancer 14:30 (2014). [cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/GB2016/052564, dated Feb. 20, 2018. [cited by applicant]
International Search Report and Written Opinion for Application No. PCT/GB2016/052564, dated Oct. 12, 2016. [cited by applicant]
Kawahara et al., “Mimicry of Erythropoietin and Interleukin-6 Signalling by an Antibody/Cytokine Receptor Chimera in Murine Myeloid 32D Cells,” J. Biochem. 141:563-571 (2007). [cited by applicant]
Kawahara, et al., “Engineering cytokine receptors to control cellular functions,” Biochemical Engineering Journal, 48:283-294 (2010). [cited by applicant]
Leen et al., “Reversal of Tumor Immune Inhibition Using a Chimeric Cytokine Receptor,” Mol Therapy, 22(6):1211-1220 (2014). [cited by applicant]
Leinonen et al., “Characterization of monoclonal antibodies against prostate specific antigen produced by genetic immunization,” J Immunol Methods, 289(1-2):157-167 (2004). [cited by applicant]
Livnah et al., “Crystallographic Evidence for Preformed Dimers of Erythropoietin Receptor Before Ligand Activation,” Science 283:987-990 (1999). [cited by applicant]
Morgenroth et al., “Targeting of tumor cells expressing the prostate stem cell antigen (PSCA) using genetically engineered T-cells,” The Prostate, 67(10):1121-1131 (2007). [cited by applicant]
Nagarkatti et al., “Constitutive activation of the interleukin 2 gene in the induction of spontaneous in vitro transformation and tumorigenicity of T cells,” PNAS, 91(16):7638-7642 (1994). [cited by applicant]
Nustad et al., “Specificity and Affinity of 26 Monoclonal Antibodies Against the CA 125 Antigen: First Report from the ISOBM TD-1 Workshop,” Tumor Biology, 17:196-219 (1996). [cited by applicant]
Shirasu et al., “Functional Design of Chimeric T-Cell Antigen Receptors for Adoptive Immunotherapy of Cancer: Architecture and Outcomes,” Anticander Research 32:2377-2384 (2012). [cited by applicant]
Sogo et al., “Selective Expansion of Genetically Modified T Cells Using an Antibody/Interleukin-2 Receptor Chimera,” J Immunol Methods, 337(1):16-23 (2008). [cited by applicant]
Stura et al., “Crystal Structure of Human Prostate-Specific Antigen in a Sandwich Antibody Complex,” J Mol Biol, 414(4):530-544 (2011). [cited by applicant]
Wilkie et al., “Selective Expansion of Chimeric Antigen Receptor-targeted T-cells with Potent Effector Function using Interleukin-4,” J Biol Chem, 285(33):25538-25545 (2010). [cited by applicant]
Argentova V.V., et al., “Study on the Influence of Different Designs of Eukaryotic Vectors on the Expression of Recombinant IgA”, Moscow University Biological Sciences Bulletin, Episode 16: Biology, 2017, vol. 72, No. 2… [cited by applicant]
Berry M.J., et al., “Substitution of Cysteine for Selenocysteine in type I Iodothyronine Deiodinase Reduces the Catalytic Efficiency of the Protein but Enhances its Translation”, Endocrinology, 1992, vol. 131, No. 4, pp… [cited by applicant]
Brunner T., et al., “Cytotoxic T cells: Double-Barreled Shot Guns,” Nature Medicine, Abstract, vol. 5, No. 1, 1999, pp. 20. [cited by applicant]
Cordoba S.P., et al., “The Large Ectodomains of CD45 and CD148 Regulate their Segregation from and Inhibition of Ligated T-cell Receptor,” Blood, May 23, 2013, vol. 121 (21), pp. 4295-4302. [cited by applicant]
Gasser B., et al., “Antibody Production with Yeasts and Filamentous Fungi: On the Road to Large Scale?”, Biotechnology letters, 2007, vol. 29, No. 2, pp. 201-212,?.208. [cited by applicant]
Jones S., et al., “Lentirival Vector Design for Optimal T Cell Receptor Gene Expression in the Transduction of Peripheral Blood Lymphocytes and Tumor-infiltrating Lymphocytes,” Human Gene Therapy, 2009, vol. 20, pp. 630… [cited by applicant]
Roitt I., et al., Immunology, Enzymatic Cleavage of IgG1, Fifth Edition, Moscow, Mir, 2000, pp. 4-6. [cited by applicant]
Sadelain M., et al., “The Basic Principles of Chimeric Antigen Receptor (CAR) Design,” Cancer Discovery, vol. 3 (4), Apr. 2013, pp. 388-398, XP055287277, doi:10.1158/2159-8290.CD-12-0548, ISSN 2159-8274. [cited by applicant]
Singer M., et al., Genes & Genomes, vol. 1 Moscow: Mir, 1, 1998, pp. 63-64. [cited by applicant]
Su C.T.-T., et al., “The role of Antibody V? Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding”, Scientific Reports, 2017, vol. 7, 3766, pp. 1-7. [cited by applicant]
Wei X-Q., et al., “The Sushi Domain of Soluble IL-15 Receptor a Is Essential for Binding IL-15 and Inhibiting Inflammatory and Allogenic Responses In Vitro and In Vivo,” The Journal of Immunology, 2001, vol. 167, No. 1,… [cited by applicant]
Yarilin A.A., “Fundamentals of Immunology,” Moscow Medicine, 1999, pp. 172-174. [cited by applicant]